Process of growing lactobacilli and the product



Patented June 9, 19 25.

um'riso srarss JOHN REIGHEL, OF PHILADELPHIA, AND CHRISTOPHER R008,

PENNSYLVANIA, ASSIGNOBS PO-H. K. MULIORD COMPANY,

PENNSYLVANIA.

PATIENT OFFICE.

or wumncronn, n oonrom'rron or PROCESS OF GROWING LACTOBACILLI AND THE PRODUCT,

Io Drawing;

To all whom it may concem:

. Be it known that we, Joan Rnrcnnn, a 4 citizen of the United States, residing Philadelphia, in the county of Philadelphia 6 and State of Pennsylvania, and CHRISTOPHER Roos, a citizen of the United States, residing at Wallingford, in the county of Delaware and State of Pennsylvania, have invented new and useful Improvements in Processes 10 of Growing Lactobacilli and the Product, of

which the following is a specification.

This invention relates to the process of growing lactobacilli, such .as Bacillrw acidoph'llus and Bacillus Bulgarz'mls in a. culture medium other than milk or whey with the object'of obtainin an increased number of bacilli in the me ium, and harvesting the v growth in a compact ingesting volume or mass and thereby producing a new and original com act form of ingestable prodnot that can be ingested in doses each containing the necessary number of bacilli in compact disipensible form and ingestable volume as 'stinguished from the voluminous doses in the quantity form of product containing the necessary number of bacilli that is dispensed and ingested as Bacillus acidophilus milk or whey.

The lactobacillu's, such as B. acidbphz'hw is an inhabitant and 8. art of the flora of the normal intestines; at it canbe made to grow outside of the intestinal tract; can be made to flourish in milk as a culture medium. In the prior stateof the art, it is familiar practice to take cultures that grow well in milk, and supply the Bacillus acido hilus for ingesting pugposes in a form of milk preparation, commo y known as B.

. acidophz'lus milk. The milk pre ara'tions 40 of B. acido hz'lus milk are prepared y growing the acidophilus organisms either in skimmed or whole milk, or whey, previously pasteurized or sterilized. The Bacillus acidophilus grown in the milk, or whey, increases rapidly in numbers, and when it reaches its maximum growth. the rescnce of lactic acid becomes noticeable, w ich is also accompanied by the thickening of he milk. The owth of the B. acidophilua is inhibited the increase in the lactic acidity which mcreases to the point of destroying or killing the bacilli in the milk culture ilt Application filed June 19, 1924. Serial No. 721,146.

has been shown that the maximum growth of B. acidoph'ilw in milk may reach .the number approximately of one billion organisms to the cubic centimeter. These figures are mentionedhere, inasmuch as' they are important in respect to the dosage of B. acidophilus milk, or B. acidophilws preparations, since it isbelieved that it is important to ingest a large number of viable B. acidopkilus, and which importance cannot be overemphasized. The therapy of lactobacillicalls for a dosage of B. acidophilus milk'of approximately one quart a day for the individual. That volume is sometimes reduced or exceeded; but it is common experience that in treating severe cases of chronic constipation it is frequently'nece'ssary to increase the dosage before obtaining beneficialresults. It is safe to say that-the customary dosage is close to one quart of B. acidophz'lus milk a day. One quart of- B. acidophz'lus milk includes nine-hundred and sixty cubic centimeters, or, in round-figures, approximately one thousand. cubic centimeters; and each cubic. centimeter of B. I

acidophilus milk with its maximum growth or bacilli will include one billion mat him bacilli. It may, therefore, be state .that

the ,average dose of viable B. acidophilus irrespective of the milk includes approximate y one thousand billion (widophz'lus bacilli. The bulk of the one thousand billion- B. aoz'dophc'lus bacilli inone quart of B. acidophz'lus milk approximates about from one to two grams in a moist state. The I volume is from one to two cubic centimeters. To supply B. acidophilus in any other medium in the dosage recommended in lactobacillus therapy, it is necessary theirefore to apy. The presentprocess isdirected to the s1ze of dose required of B. acidophiI/ae for include approximately one thousandblllionthe average person; namely, the number of B. aculo hilus organisms in one quart of B. aa'glophz as milk, one thousand billion am'dophiI/us-baoilli} with the object of reducing the volume of the media containing the bacilli, but nevertheless retaining the necessary number of bacilli in the dose, as referred to above.

In the present invention it has been demonstrated that a culture media other than skimmed or whole milk or whey can be prepared at a cost not exceeding the cost of milk, with growth factors al owing a rapid growth of B. am'dophitus to a degree of concentration of number of viable B.. aoidophz'lus that will permit of the establishmentof thedose in a small volume with the necessary number of bacilli as compared with a doseof B. 'am'dophilus milk. The medium used for the purpose of growing B. aoz'doph'il'us or B. Bulgarious in the present invention is prepared by digesting milk or casein with a ferment or trypsin in the proportion based on the tryptic value of the ferment. It 'is rendered alkaline with sodium carbonate, and incubated six hours at. 42 to 45 C. The medium is diluted with three to four volumes of water and the reactionadjusted to PH 7.0. The medium is sterilized under substantially fifteen pounds pressure for a proximately thirty'minutes in glass lined igestors, then cooled to about 40 (3., and planted with B. zwz'dophilus or B. Bulga'ric'us seed. The special advantages obtained by growing the bacilli in the artificial medium prepared by digesting milk or casein with a ferment or trypsin in the preparation based on the tryptic value of the ferment, especially as -'com ared with the advantages of growing the bacilli in a culture medium of glucose broth, or glucose agar, are as follows: It ossesses growth factors which encourage t e rapid growth of viable bacilli in higher concentration than milk or whey, their growth being several times that attained or obtained in milk or whey, and the growth as a whole comprises a lactobacillus mass.

In carr ing out the process a pure culture of B. ophihw or B. Bulgaricus'is taken, and regarded asthe seed. The temperature is maintained at about 40 C. for approximately thirty hours or until the maximum growth is obtained. 'The growth is removed from the culture media or harvested by means of settling, or by the use of the centrifu e. The mass of B. amidophi- Zus or B. Bu wr'icus, in each two grams, represents approximately the growth of B. acidophilus equal to the number of one thousand billion B. aoidophz'lua obtained in a quart of B. am'dopkz'ms milk. The harvested mass of lactobacilli in the proportion stated, namely two grams, can be used in a moist state, with the addition of a filler, such'as tapioca, flour, starch, sugar, gelatin, or other suitable-substance, be compressed, or it can be used without a filler,

or it can be mixed with yeast, and subsequently be compressed into cakes, pellets, tablets, cylinders, or other forms, or the mass can be dried to a powder, or granular condition, and be converted into tablets. It is understood that the present invention is not limited to the use of the s ecific culture media defined herein in describing the process, but that the invention may include any other artificial culture media, or any natural culture media other than milk, skimmed milk' or whole milk or whey, that is favorable forthe growth of lactobacilli ,of an increased number with the object of attaining the present invention, which is the production of lactobacilli in plastic mass in a readily ingestible form whereby a convenient dose of lactobacilli may be obtained containing the approximate number of lactobacilli in a quart of lactobacillus milk. The culture medias intended to be used in the invention include such culture medias as will permit of the collection of the bacilli from the culturemedia for the purpose of harvesting or gathering them, in order to obtain several or many times the required dose of lactobacilli in a small ingestible mass or volume as compared with the dose of lactobacillus milk. Thus, it will be perceived that the required dose of B. acidophihas in the present invention isin a more extended useful form than thatproduced in B. acidop hilus milk, and among other matters that may be stated, the product is obtained in smaller volume or mass than in lactobacillus milk; and the product vis of signal utility asenabling the ingesting of the required number of bacilli in a dose without having to ingest as. well a lar volume of culture media milk, or ingest mi k at all, as a part of the dose of bacilli, in order to ingest the requirednumber of B. am'dophz'lus or B. Bulgam'cus; and further, it is of large utility in the art of producing the products of lactobacillus, such as ,B acidoph'ihze and B. Bulgam'ous in a culture media other than milk, for putting the prodnot on sale, and dispensing" it, on a large scale, as the handlin treating, shipping, storing, dispensing an replacement of large quantities of milk is avoided; in brief, the Y ing and enlarging the use of lactobacilli in i a culture media other than milk or whey commonly by personswhose health mi ht be materially enefitted could they ingest B. aoz'dophihaa or. B; Bulgm'irms in the pre-' scribed proper number in accordance with the prescribed rules of lactobacillus therapy.

"mass con" state, with the ophz'lua milk,

It was written above that the harvested mass of lactobacilli can be used in a moist addition of a filler; it is found that gelatin,

only serves to hold the bacilli together in masspbutas it fills the interstices between the bacilli and also occludes themas a'mass, it serves to preserve or retain the. moisture or moist state of the bacilli, which is a valu-- able function, since only the Viable organisms have lactobacillus therapeutic quality, and, therefore, if the moisture dries out the viability of the bacilli is greatly depleted, if not entirely lost. The use of gelatin therefore with the bacilli enables theprodnet to remain etlicient for a long time in respect to placin it on sale commercially for dispensing, an does not require the stock of bacilli on sale for dispensing purposes to be frequently replaced with new or fresh stock. In the case of B. acidophil-us milk themilk kept the bacilli in a moist state, but that advantage was greatly lessened in commercially dispensi g the product because of the fact that. B. acidophiluc milk on sale for dispensing soon becomes stale, and has to be re laced with fresh 'B acids opkz'lua milk. The gelatine occlude product can be sugar coated for the palate if desired. In mentioning gelatin, we do not intend to exclude any vehicle that can be used fora similar purpose.

Having thus fully described our invention, whatwe claim as new and desire to secure by Letters Patent is: r 1. A method for obtaining lactobacillus be mass contaming substantially the same number of lactobacilli containedin B. acidor Bulgariouamilk, which consists in growing lactobacilli in a culture media or other than milk or whey.

2. A method of obtaining lactobacilli substantially the same number of lacto acilli contained in milk or B. Bulgzmims milk, which consists ingrowmg lactobacilli in a culture media preparediby digesting casein with a'ferment in proportions smted to the purpose treating the mixture with an alkali, incubating the same for substantially shapes containing substantially the same number of lactobacilli contained in lacto' bacillus milk, which consists in lactobacilli an artificial culture or like substances, is highly suitable for the purpose, since it not" vqulred number of in a culture cubic cen culture 'lactobacilll in six hours at 45 C. star-- v lhflmg, and cooling substantially to 40 (3.

edia,

ing it yeast, and compressing the mix- .ture into desired forms.

4.A method for obtaining lactobacilli medium with try 'ilsin and then with an alkali, incubatin e same for substantially six hours at 45 6., sterilizing, cooling substantially to 40" media with lactobacillu and allowing them sufiicient time to attain their. maximum growth,

edi

removing the growth from the m a i 5 A method of obtaining lactobacillus mass with yeast and compressed into desired forms containing substantially same concentration of lactobacilli contained. in lactobacillus milk, which consists in ad?- ing to a culture medium a ferment capabe of adding to the growth factors therein, planting the medium with lactobacillus, allowing them suflicient time to attain their maxnnnm growth, removing the growth from the medium, and mixing it with yeast and compressing the mixture into desir forms. p

6. A new article of production, comprising a lactobacillus mass, containing the re- B. acidopkibus to the dose of lactobacillus milk and a culture media of smaller volume than lactobacillus milk.

7. A new media other than milk, thelactobacilli being approximately one to two timeters or two grams, the same in num r of-B. lactobacillus milk.

8. A new article of production, comprising the mitted the roduction of lactobacilli insubstantially the same number the dose as that produced in tobacillus V -9-.A new article of ing the required dose 0 lactobac' in a cul ture media that permitted the productionv substantially the same num hh'er in the required dose as thatproduced e the culture media. of the required dose is of less volume than'the milk media contained in said required dose; i

10. A new article of production, comprised of lactobacillus mass in a moist state and gelatin. 7.

' Signed this 17th day of June, 1994.

- JOHN REICHEL.

CHRISTOPHER ROOS.

the required dose of lactheproduct comprising lactobacilli 1n concentration not less than the us in one quart of fproduction comprisrequired'dose of lactobacillus) milk, but "115 O. and then planting the retgxiired doseof lactobacilli m a me 'a other than milk that per- 

